Phosphopeptide mapping of plexiform lesions in pulmonary arterial hypertension

肺动脉高压丛状病变的磷酸肽图谱

基本信息

批准号：
8211870
负责人：
LAWRENCE S. ZISMAN
金额：
$ 7.25万
依托单位：
PULMOKINE, INC.
依托单位国家：
美国
项目类别：
财政年份：
2011
资助国家：
美国
起止时间：
2011-09-30 至 2013-08-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8211870
关键词：
Address Algorithms Bioinformatics Blood Vessels Blood flow Cells Chromatography Clinical Trials Data Disease Family Freezing Frozen Sections Goals Growth Imatinib Immunoassay Incubated Intervention Knowledge Label Lead Lesion Lung Maps Mass Spectrum Analysis Methods Morbidity - disease rate Output Pathology Patients Peptides Phase Phosphopeptides Phosphoproteins Phosphorylation Phosphorylation Site Phosphotransferases Proliferating Protein Analysis Protein Isoforms Protein-Serine-Threonine Kinases Proteins Proteomics Protocols documentation Public Health Rare Diseases Receptor Protein-Tyrosine Kinases Regulation Relative (related person)Research Role Sampling Societies Structure of parenchyma of lung Techniques Testing Tissues Tyrosine Tyrosine Kinase Inhibitor abstracting arteriole blood lead candidate identification cell growth experience hypertension control innovation interest kinase inhibitor laser capture microdissection mortality nanofluidic new therapeutic target novel oncology protein aminoacid sequence pulmonary arterial hypertension therapeutic target titanium dioxide validation studies

项目摘要

DESCRIPTION (provided by applicant): Pulmonary Arterial Hypertension (PAH) is a rare disorder of the pulmonary vasculature associated with high morbidity and mortality. The pathology of the disease consists of plexiform lesions of proliferating cells which obstruct blood flow through the pulmonary arterioles. There is a growing interest in the use of kinase inhibitors to address this underlying pathology. However, very little is actually understood about the regulation of kinases in PAH, and the phosphoproteome in PAH has not yet been studied. This project will determine the phosphopeptide profile of lung tissue from subjects with idiopathic pulmonary arterial hypertension (iPAH) compared to controls. Titanium dioxide chromatography will be used to enrich for phosphopeptides which will then be identified by mass spectroscopy. A bioinformatics approach will be used to predict the kinases and/or kinase families most likely to be responsible for these increased phosphoproteins. The NetworKIN and other algorithms will be used for this purpose. The predictions of NetworkKIN will be tested in a subset of kinases by incubating the selected recombinantly expressed kinases with peptide microarrays that duplicate the peptide sequences of the candidate phosphorylation sites. The findings of the proteomic analysis will be validated by isolating protein from plexiform lesions with laser capture microdissection. The proteins will be analyzed with a novel nanofluidic immunoassay that can detect very small quantities of protein. The results of this study could identify new therapeutic targets for the treatment of iPAH and thereby benefit patients and society. PUBLIC HEALTH RELEVANCE: Pulmonary Arterial Hypertension (PAH) is a rare disorder of the pulmonary blood vessels associated with high morbidity and mortality. The purpose of this project is to identify proteins that may be responsible for the abnormal growth of cells that block the blood vessels and lead to the disease. This project is relevant to public health because it could lead to new treatments for PAH. (End of Abstract)

描述（由申请人提供）：肺动脉高压（PAH）是一种罕见的肺血管疾病，具有高发病率和死亡率。该疾病的病理学包括增殖细胞的丛状病变，其阻碍通过肺小动脉的血流。人们越来越关注使用激酶抑制剂来解决这种潜在的病理学问题。然而，人们对 PAH 中激酶的调控实际上知之甚少，并且 PAH 中的磷酸蛋白质组尚未得到研究。该项目将确定特发性肺动脉高压 (iPAH) 受试者与对照组相比肺组织的磷酸肽谱。二氧化钛色谱法将用于富集磷酸肽，然后通过质谱法进行鉴定。将使用生物信息学方法来预测最有可能导致这些磷蛋白增加的激酶和/或激酶家族。 NetwoKIN 和其他算法将用于此目的。 NetworkKIN 的预测将在激酶子集中进行测试，方法是将选定的重组表达激酶与复制候选磷酸化位点肽序列的肽微阵列一起孵育。蛋白质组分析的结果将通过激光捕获显微切割从丛状病变中分离蛋白质来验证。这些蛋白质将通过一种新型纳米流体免疫测定法进行分析，该免疫测定法可以检测极少量的蛋白质。这项研究的结果可以确定治疗 iPAH 的新治疗靶点，从而造福患者和社会。公共卫生相关性：肺动脉高压 (PAH) 是一种罕见的肺血管疾病，发病率和死亡率较高。该项目的目的是确定可能导致细胞异常生长的蛋白质，从而阻塞血管并导致疾病。该项目与公共卫生相关，因为它可能会带来针对多环芳烃的新疗法。（摘要完）